The present invention relates to fused heteroaryl derivatives which are useful as medicaments, more particularly as phosphatidylinositol 3-kinase (PI3K) inhibitors and carcinostatic agents.
Phosphatidylinositol (hereinafter abbreviated as xe2x80x9cPIxe2x80x9d) is one of phospholipids in cell membranes. In recent years it has become clear that PI plays an important role also in intracellular signal transduction. It is well recognized in the art that especially PI (4,5) bisphosphate (PI(4,5)P2) is degraded into diacylglycerol and inositol (1,4,5) triphosphate by phospholipase C to induce activation of protein kinase C and intracellular calcium mobilization, respectively [M. J. Berridge et al., Nature, 312, 315 (1984); Y Nishizuka, Science, 225, 1365 (1984)].
Turning back to the late 1980s, PI3K was found to be an enzyme to phosphorylate the 3-position of the inositol ring of phosphatidylinositol [D. Whitman et al., Nature, 332, 664 (1988)].
PI3K was originally considered to be a single enzyme at the time when PI3K was discovered. Recently it was clarified that a plurality of subtypes are present in the PI3K. Three major classes of PI3Ks have now been identified on the basis of their in vitro substrate specificity [B. Vanhaesebroeck, Trend in Biol. Sci., 22, 267(1997)].
Substrates for class I PI3Ks are PI, PI(4)P and PI(4,5)P2. In these substrates, PI(4,5)P2 is the most advantageous substrate in cells. Class I PI3Ks are further divided into two groups, class Ia and class Ib, in terms of their activation mechanism. Class Ia PI3Ks, which include PI3K p110xcex1, p110xcex2 and p110xcex4 subtypes, are activated in the tyrosine kinase system. Class Ib PI3K is a p110xcex3 subtype activated by a G protein-coupled receptor.
PI and PI(4)P are known as substrates for class II PI3Ks but PI(4,5)P2 is not a substrate for the enzymes of this class. Class II PI3Ks include PI3K C2xcex1, C2xcex2 and C2xcex3 subtypes, which are characterized by containing C2 domains at the C terminus, implying that their activity will be regulated by calcium ions. The substrate for class III PI3Ks is PI only. A mechanism for activation of the class III PI3Ks is not clarified yet. Since each subtype has its own mechanism for the regulating activity, it is considered that the respective subtypes will be activated depending on their respective stimuli specific to each of them.
In the PI3K subtypes, the class Ia subtype has been most extensively investigated to date. The three subtypes of class Ia are hetero dimers of a catalytic 110 kDa subunit and regulatory subunits of 85 kDa and 55 kDa. The regulatory subunits contain SH2 domains and bind to tyrosine residues phosphorylated by growth factor receptors with a tyrosine kinase activity or oncogene products thereby inducing the PI3K activity of the p110 catalytic subunit. Thus, the class Ia subtypes are considered to be associated with cell proliferation and carcinogenesis. Furthermore, the class Ia PI3K subtypes bind to activated ras oncogene to express their enzyme activity. It has been confirmed that the activated ras oncogene is found to be present in many cancers, suggesting a role of class Ia PI3Ks in carcinogenesis.
As explained above, PI3K inhibitors are expected to be a novel type of medicaments useful against cell proliferation disorders, especially as carcinostatic agents. As for the PI3K inhibitor, wortmannin [H. Yano et al., J. Biol. Chem., 263, 16178 (1993)] and LY294002 [J. Vlahos et al., J. Biol. Chem., 269, 5241(1994)] which is represented by the formula below are known. However, development of PI3K inhibitors having a more potent cancer cell growth inhibiting activity is desired. 
Japanese Patent KOKAI (Laid-Open) No. 6-220059 discloses fused heteroaryl derivatives shown by formula (a) below which possess an activity of reducing the blood glucose level. Furthermore, compounds shown by formula (b) and formula (c) below are described in Indian J. Chem., Sect. B (1993), 32B (9), 965-8 and J. Heterocycl. Chem. (1992), 29 (7), 1693-702, respectively. In addition, Al-AzharBull. Sci. (1992), 3(2), 767-75 discloses a compound shown by formula (d) below. However, none of these prior art publications disclose or suggest the PI3K inhibiting activity. 
In formula (a) above, Z is O, S or xe2x95x90Nxe2x80x94R0, R1 is an amino which may be substituted, a heterocyclic group which may be substituted, etc.; R2 is cyano, an amino which may be substituted, or a heterocyclic group which may be substituted; and with respect to the remaining substituents, see the specification of the patent. In formula (b) and (c) above, R is a (substituted) amino or a (substituted) nitrogen-containing saturated heterocyclic group.
Publication No. WO98/23613 discloses fused pyrimidine derivatives, such as 7H-pyrrolo[2,3-d]pyrimidine derivatives, which having a tyrosine kinase receptor inhibiting activity and which are useful as carcinostatic agents, wherein the fused pyrimidine derivatives have at its fourth position a particular-heteroaryl-substituted amino, pheny-substituted amino, or indole-1-yl, and have no substituent at its second position.
Following compounds are known among the compounds shown by general formula (I), whereas xe2x80x9cAxe2x80x9d ring is a ring shown by (b);
(1) Ann. Pharm. Fr. (1974), 32(11), 575-9 discloses 4-(4-morpholinyl)-2-phenylpirido[2,3-d]pyrimidine as a compound having antiinflammatory and spasmolytic activities,
(2) Chem. Pharm. Bull. (1976), 24(9), 2057-77 discloses 4-(4-morpholinyl)-2-phenylpirido[2,3-d]pyrimidine-7(1H)-one as a compound having a diuretic activity,
(3) Khim.-Farm. Zh. (1993), 7(7), 16-19 and Khim. Geterotsiki. Soedin. (1971), 7(3), 418-20 disclose 4-(4-morpholinyl)-2-phenyl-6-quinazolinol and 6-methoxy-4-(4-morpholinyl)-2-phenylquinazoline as compounds having an antibiotic activity,
(4) Publication No. WO2000/41697 discloses 2,4-diamino-6-phenyl-8-piperidinopyrimido[5,4-d]pyrimidine as a compound having celebral ischemia prevention and treatment effects,
(5) Publication No. WO99/32460 discloses, as cardiovascular drugs, compounds of general formula (Ib) described hereinafter wherein B is a benzene ring, W is N, n is 2 or 3, existing R1""s are all xe2x80x94OMe, and R4b is an unsubstituted phenyl or a phenyl substituted by 1 to 3 substituents which are selected from -a halogen, NO2, -a lower alkyl, xe2x80x94O-a lower alkyl, -a halogenated lower alkyl and xe2x80x94CONRaRc,
(6) Publication No. BE841669 discloses, as antiparasitics, compounds of general formula (Ib) described hereinafter wherein B is a benzene ring, W is N, n is 1, R1 is -a halogen or -a lower alkyl, and R4b is -(an imidazolyl which may have one or more substituents),
(7) Publication No. WO99/43682 discloses, as antianxiety agents, compounds of general formula (Ib) described hereinafter wherein B is a thiophene ring, and W is CH,
(8) Japanese Patents KOKAI (Laid-Open) Nos. 62-10085 and 61-158983 disclose compounds of general formula (Ib) described hereinafter wherein B is an imidazole ring, and W is N, whereas the compounds have an antiinflammatory activity, a platelet aggregation inhibiting activity, etc.,
(9) U.S. Pat. No. 3,873,545 and Act Pol. Pharm. (1994), 51(4-5), 359-63 disclose compounds of general formula (Ib) described hereinafter wherein B is a pyridine ring, and R4b is an unsubstituted phenyl, an unsubstituted pyridyl, or -a lower alkylene-(a nitrogen-containing saturated heterocyclic group which may have one or more substituents), whereas the compounds have a spasmolytic, diuretic or hypotensive activity,
(10) U.S. Pat. No. 2,940,972 discloses compounds of general formula (Ib) described hereinafter wherein B is a pyrazine ring, and R4b is an unsubstituted phenyl, or a benzyl, whereas the compounds have a coronary dilating or sedative activity,
(11) U.S. Pat. No. 3,753,981 and Germnan Patent Publication No. 2,140,280 disclose compounds of general formula (Ib) described hereinafter wherein B is a benzene ring, and R4b is a styryl or 2-(5-nitro-2-furyl)vinyl, whereas the compounds have an antiinflammatory or antibiotic activity, and
(12) Eur. J. Med. Chem. (1996), 31(5), 417-425, discloses compounds of general formula (Ib) described hereinafter wherein B is a benzene ring, W is CH, and R2 and R3 are bonded together with an adjacent N atom to form -(piperidinyl which may have one or more substituents) or -(piperazinyl which may have one or more substituents), as compounds working as a benzodiazepine receptor ligand, U.S. Pat. No. 4,560,692 discloses them as those having a spasmolytic and ataractic activity, and Japanese Patents KOKAI (Laid-Open) No. 2-129169 discloses them as those having a lipoperoxidation inhibiting activity.
Furthermore, compounds of general formula (Ib) described hereinafter wherein B is a pyridine ring and n is 0 are disclosed in Japanese Patent KOKAI (Laid-Open) No. 51-138689 (antiparasitics), Japanese Patent KOKAI (Laid-Open) No. 56-120768 (a dye component for thermosensitive recording materials), Antimicrob. Agents Chemother., (1975), 8 (2), 216-19 (an antibacterial activity), Cancer Res. (1975), 35 (12), 3611-17 (a mutagenic activity), CA 64: 19608c, Collect. Czech. Chem. Commun., (1994), 59 (6), 1463-6, U.S. Pat. No. 5,304,554 (an anti-HIV activity), Chem. Pharm. Bull., (1982), 30(6), 1974-9, and J. Heterocycl. Chem. (1980), 17(5), 1029-34. However, none of the prior publications teach or even suggest the PI3K inhibiting activity and carcinostatic activity.
The present inventors have performed extensive investigations on compounds with a PI3K inhibiting activity. As a result, it has been found that novel fused heteroaryl derivatives have an excellent PI3K inhibiting activity as well as a cancer cell growth inhibiting activity. Based on the finding, it has been discovered that the fused heteroaryl derivatives could be excellent PI3K inhibitors and antitumor agents. The present invention has thus been achieved.
Therefore, the present invention relates to pharmaceutical compositions, which are PI3K inhibitors or antitumor agents, comprising a fused heteroaryl derivative represented by general formula (I) below or a salt thereof and a pharmaceutically acceptable carrier. 
wherein: 
B represents a benzene ring, or a 5- or 6-membered monocyclic heteroaryl containing 1 to 2 hetero atoms selected from O, S and N;
R1 represents -a lower alkyl, -a lower alkenyl, -a lower alkynyl, -a cycloalkyl, -an aryl which may have one or more substituents, -a heteroaryl which may have one or more substituents, -a halogen, xe2x80x94NO2, xe2x80x94CN, -a halogenated lower alkyl, xe2x80x94ORb, xe2x80x94SRb, xe2x80x94SO2-Rb, xe2x80x94SOxe2x80x94 Rb, xe2x80x94COORb, xe2x80x94COxe2x80x94Rb, xe2x80x94CONRaRb, xe2x80x94SO2NRaRb, xe2x80x94NRaRb, xe2x80x94NRaxe2x80x94CORb, xe2x80x94NRaxe2x80x94SO2Rb, xe2x80x94Oxe2x80x94 COxe2x80x94NRaRb or xe2x80x94NRaCOxe2x80x94COORb, xe2x80x94CO-a nitrogen-containing saturated heterocyclic group, xe2x80x94CONRa-a lower alkylene-ORb, xe2x80x94CONRa-a lower alkylene-NRb, xe2x80x94O-a lower alkylene-ORb, xe2x80x94O-a lower alkylene-O-a lower alkylene-ORb, xe2x80x94O-a lower alkylene-NRaRb, xe2x80x94O-a lower alkylene-O-a lower alkylene-NRaRb, xe2x80x94O-a lower alkylene-NRc-a lower alkylene-NRaRb, xe2x80x94NRc-a lower alkylene- NRaRb, xe2x80x94N(a lower alkylene-NRaRb)2, xe2x80x94CONRaxe2x80x94ORb, xe2x80x94NRaxe2x80x94COxe2x80x94NRbRc, or xe2x80x94OCORb;
each of R2 and R3, which may be the same or different, represents xe2x80x94H, -a lower alkyl, -a lower alkylene-ORa or -a lower alkylene-NRaRc, or R2 and R3 are combined together with the N atom adjacent thereto to form a nitrogen-containing saturated heterocyclic group as xe2x80x94NR2R3 which may have one or more substituents;
each of Ra and Rc, which may be the same or different, represents xe2x80x94H or -a lower alkyl;
Rb represents xe2x80x94H, -a lower alkyl, a cycloalkyl, an aryl which may have one or more substituents or a heteroaryl which may have one or more substituents;
n represents 0, 1, 2 or 3;
each of W and X, which may be same or different, represents N or CH;
Y represents O, S or NH; and,
R4 represents xe2x80x94H, -a lower alkyl, -a lower alkenyl, -a lower alkynyl, -(an aryl which may have one or more substituents), -a lower alkylene-(an aryl which may have one or more substituents), -a lower alkenylene-(an aryl which may have one or more substituents), -a lower alkynylene-(an aryl which may have one or more substituents), -(a cycloalkyl which may have one or more substituents), -(a cycloalkenyl which may have one or more substituents), -a lower alkylene-(a cycloalkyl which may have one or more substituents), -a lower alkenylene-(a cycloalkyl which may have one or more substituents), -a lower alkylene-(a nitrogen-containing saturated heterocyclic group which may have one or more substituents), -a lower alkenylene-(a nitrogen-containing saturated heterocyclic group which may have one or more substituents), -(a heteroaryl which may have one or more substituents), -a lower alkylene-(a heteroaryl which may have one or more substituents), or -a lower alkenylene-(a heteroaryl which may have one or more substituents). The same applies hereinbelow.
The compounds (I) of the present invention encompass the known compounds as well as commercially available compounds later described in Compound Z, which are all included within the definition of formula (I).
The present invention further relates to a novel fused heteroaryl derivative represented by general formula (Ia) or (Ib) or salts thereof, as well as a novel pharmaceutical composition comprising the same and a pharmaceutically acceptable carrier: 
[wherein:
R1 represents -a lower alkyl, -a lower alkenyl, -a lower alkynyl, -a cycloalkyl, -an aryl which may have one or more substituents, -a heteroaryl which may have one or more substituents, -a halogen, xe2x80x94NO2, xe2x80x94CN, -a halogenated lower alkyl, xe2x80x94ORb, xe2x80x94SRb, xe2x80x94SO2-Rb, xe2x80x94SOxe2x80x94 Rb, xe2x80x94COORb, xe2x80x94COxe2x80x94Rb, xe2x80x94CONRaRb, xe2x80x94SO2NRaRb, xe2x80x94NRaRb, xe2x80x94NRaxe2x80x94CORb, xe2x80x94NRaxe2x80x94SO2Rb, xe2x80x94Oxe2x80x94 COxe2x80x94NRaRb or xe2x80x94NRaCOxe2x80x94COORb, xe2x80x94CO-a nitrogen-containing saturated heterocyclic group, xe2x80x94CONRa-a lower alkyl-ORb, xe2x80x94CONRa-a lower alkylene-ORb, xe2x80x94O-a lower alkylene-NRb, xe2x80x94O-a lower alkylene-O-a lower alkylene-ORb, xe2x80x94O-a lower alkylene-NRaRb, xe2x80x94O-a lower alkylene-O-a lower alkylene-NRaRb, xe2x80x94O-a lower alkylene-NRc-a lower alkylene-NRaRb, xe2x80x94NRc-a lower alkylene-NRaRb, xe2x80x94N(a lower alkylene-NRaRb)2, xe2x80x94CONRaxe2x80x94NRb, xe2x80x94NRaxe2x80x94COxe2x80x94NRbRc, or xe2x80x94OCORb;
each of R2 and R3, which may be the same or different, represents xe2x80x94H or -a lower alkyl, or R2 and R3 are combined together with the N atom adjacent thereto to form a nitrogen-containing saturated heterocyclic group as xe2x80x94NR2R3 which may have one or more substituents;
Ra and Rc, which may be the same or different, represent xe2x80x94H or -a lower alkyl;
Rb represents xe2x80x94H, -a lower alkyl, a cycloalkyl, an aryl which may have one or more substituents or a heteroaryl which may have one or more substituents;
n represents 0, 1, 2 or 3;
X represents N or CH;
Y represents O, S or NH; and,
R4a represents -(an aryl which may have one or more substituents), -a lower alkylene-(an aryl which may have one or more substituents), -a lower alkenylene-(an aryl which may have one or more substituents), -a lower alkynylene-(an aryl which may have one or more substituents), -(a cycloalkyl which may have one or more substituents), -(a cycloalkenyl which may have one or more substituents), -a lower alkylene-(a cycloalkyl which may have one or more substituents), -a lower alkenylene-(a cycloalkyl which may have one or more substituents), -a lower alkylene-(a nitrogen-containing saturated heterocyclic group which may have one or more substituents), -a lower alkenylene-(a nitrogen-containing saturated heterocyclic group which may have one or more substituents), -(a heteroaryl which may have one or more substituents), -a lower alkylene-(a heteroaryl which may have one or more substituents), or -a lower alkenylene-(a heteroaryl which may have one or more substituents);
with the proviso that the following compounds are excluded:
(1) compounds in which X represents N, Y represents S, n is 3 and R1 represents a combination of xe2x80x94CN, xe2x80x94OEt and phenyl, and R4a represents 2-nitrophenyl;
(2) compounds in which X represents CH, and R4a represents -(a heteroaryl which may have one or more substituents);
(3) compounds in which X represents CH, Y represents O, n is 0 and R4a represents an unsubstituted phenyl; and
(4) compounds in which X represents N, Y represents S, n is 2, R1 represents an unsubstituted phenyl and R4a represents 4-methoxyphenyl or an unsubstituted phenyl. The same applies hereinbelow. 
[wherein:
B represents a benzene ring, or a 5- or 6-membered monocyclic heteroaryl containing 1 to 2 hetero atoms selected from O, S and N;
R1 represents -a lower alkyl, -a lower alkenyl, -a lower alkynyl, -a cycloalkyl, -an aryl which may have one or more substituents, -a heteroaryl which may have one or more substituents, -a halogen, xe2x80x94NO2, xe2x80x94CN, -a halogenated lower alkyl, xe2x80x94ORb, xe2x80x94SRb, xe2x80x94SO2-Rb, xe2x80x94SOxe2x80x94 Rb, xe2x80x94COORb, xe2x80x94COxe2x80x94Rb, xe2x80x94CONRaRb, xe2x80x94SO2NRaRb, xe2x80x94NRaRb, xe2x80x94NRaxe2x80x94CORb, xe2x80x94NRaxe2x80x94SO2Rb, xe2x80x94Oxe2x80x94 COxe2x80x94NRaRb, xe2x80x94NRaCOxe2x80x94COORb, xe2x80x94NRaCOORb, xe2x80x94NRaCO-a lower alkylene-an aryl, xe2x80x94NRaxe2x80x94SO2- a lower alkylene-an aryl, xe2x80x94NRa-a lower alkylene-an aryl, -a lower alkylene-ORb, -a lower alkylene-NRaRb, xe2x80x94CO-a nitrogen-containing saturated heterocyclic group, xe2x80x94CONRa-a lower alkylene-ORb, xe2x80x94CONRa-a lower alkylene-NRcRb, xe2x80x94CONRa-a lower alkylene-a nitrogen- containing saturated heterocyclic group, xe2x80x94O-a lower alkylene-ORb, xe2x80x94O-a lower alkylene- NRaRb, xe2x80x94O-a lower alkylene-a nitrogen-containing saturated heterocyclic group, xe2x80x94O-a lower alkylene-O-a lower alkylene-ORb, xe2x80x94O-a lower alkylene-O-a lower alkylene-NRaRb, xe2x80x94O-a lower alkylene-NRc-a lower alkylene-NRaRb, xe2x80x94NRc-a lower alkylene-NRaRb, xe2x80x94N(a lower alkylene-NRaRb)2, xe2x80x94CONRaxe2x80x94ORb, xe2x80x94NRaxe2x80x94COxe2x80x94NRbRc, or xe2x80x94OCORb;
R2 and R3 are combined together with the N atom adjacent thereto to form xe2x80x94NR2R3 which is a nitrogen-containing saturated heterocyclic group which may have one or more substituents;
Ra and Rc, which may be the same or different, represent xe2x80x94H or -a lower alkyl;
Rb represents xe2x80x94H, -a lower alkyl, -a cycloalkyl, -(an aryl which may have one or more substituents) or -(a heteroaryl which may have one or more substituents);
n represents 0, 1, 2 or 3, whereas n represents 1, 2 or 3 when B represents a benzene ring;
W represents N or CH; and,
R4b represents-(an aryl which may have one or more substituents), -a lower alkylene-(an aryl which may have one or more substituents), -a lower alkenylene-(an aryl which may have one or more substituents), -a lower alkynylene-(an aryl which may have one or more substituents), -(a cycloalkyl which may have one or more substituents), -(a cycloalkenyl which may have one or more substituents), -a lower alkylene-(a cycloalkyl which may have one or more substituents), -a lower alkenylene-(a cycloalkyl which may have one or more substituents), -a lower alkylene-(a nitrogen-containing saturated heterocyclic group which may have one or more substituents), -a lower alkenylene-(a nitrogen-containing saturated heterocyclic group which may have one or more substituents), -(a heteroaryl which may have one or more substituents), -a lower alkylene-(a heteroaryl which may have one or more substituents), or -a lower alkenylene-(a heteroaryl which may have one or more substituents);
with the proviso that the following compounds are excluded:
(1) 4-(4-morpholinyl)-2-phenylpyrido[2,3-d]pyrimidine,
(2) 4-(4-morpholinyl)-2-phenylpyrido[2,3-d]pyrimidin-7(1H)-one,
(3) 4-(4-morpholinyl)-2-pheny-6-quinazolinol and 6-methoxy-4-(4-morpholinyl)-2-phenyquinazoline,
(4) 2,4-diamino-6-phenyl-8-piperidinopyrimido[5,4-d]pyrimidine,
(5) compounds in which B represents a benzene ring, W represents N, n is 2 or 3, existing R1""s all represent xe2x80x94OMe, and R4b is an unsubstituted phenyl or a phenyl which is substituted by 1 to 3 substituents selected from -halogen, xe2x80x94NO2, -a lower alkyl, xe2x80x94O-a lower alkyl, -a hanogenated lower alkyl and xe2x80x94CONRaRc,
(6) compounds in which B represents a benzene ring, W represents N, n is 1, R1 represents -halogen or -a lower alkyl, and R4b represents -(imidazolyl whch may have one or more substituents),
(7) compounds in which B represents a thiophene ring, and W represents CH,
(8) compounds in which B represents an imidazole ring, and W represents N,
(9) compounds in which B represents a pyridine ring, and R4b represents an unsubstituted phenyl, an unsubstituted pyridyl, or -a lower alkylene-(a nitrogen-containing saturated heterocyclic group which may have one or more substituents),
(10) compounds in which B represents a pyrazine ring, and R4b represents an unsubstituted phenyl, or a benzyl,
(11) compounds in which B represents a benzene ring, and R4b represents a styryl or 2-(5-nitro-2-furyl)vinyl, and
(12) compounds in which B represents a benzene ring, W represents CH, and R2 and R3 are combined together with the N atom adjacent thereto to form -(piperidinyl which may have one or more substituents) or -(piperazinyl which may have one or more substituents).
The same applies hereinbelow.
Further teaching of the present invention provides a method to treat disorders (especially cancers) which are associated with PI3K, wherein the method comprises of administering to a patient an effective amount of a fused heteroaryl derivative of formula (I), (Ia) or (Ib) above or a salt thereof as well as a use of said fused heteroaryl derivative or a salt thereof for producing a medicament (especially a carcinostatic agent) which inhibit PI3K.
The compounds of general formula (I), (Ia) or (Ib) are described below in more detail.
The term xe2x80x9clowerxe2x80x9d throughout the specification is used to mean a straight or branched hydrocarbon chain having 1 to 10, preferably 1 to 6, and more preferably 1 to 3 carbon atoms.
Preferred examples of the xe2x80x9clower alkylxe2x80x9d are an alkyl having 1 to 3 carbon atoms, more preferably methyl and ethyl. Preferred examples of the xe2x80x9clower alkenylxe2x80x9d include vinyl, allyl, 1-propenyl, isopropenyl, 1-butenyl, 2-butenyl and 3-butenyl. Preferred examples of the xe2x80x9clower alkynylxe2x80x9d include ethynyl, 1-propynyl, 2-propynyl, 1-butynyl, 2-butynyl, 3-butynyl and 1-methyl-2-propynyl. The terms xe2x80x9clower alkylenexe2x80x9d, xe2x80x9clower alkenylenexe2x80x9d and xe2x80x9clower alkynylenexe2x80x9d are used to mean bivalent groups of the lower alkyl, lower alkenyl and lower alkynyl described above. Preferred examples of these groups are methylene, ethylene, vinylene, propenylene, ethynylene and propynylene. The terms xe2x80x9ccycloalkylxe2x80x9d and xe2x80x9ccycloalkenylxe2x80x9d refer to cycloalkyl and cycloalkenyl groups preferably having 3 to 8 carbon atoms. Preferred examples of these groups include cyclopropyl, cyclopentyl, cyclohexyl and cyclopentenyl.
Examples of the xe2x80x9chalogenxe2x80x9d are F, Cl, Br and I. Examples of the xe2x80x9chalogenated lower alkylxe2x80x9d are the aforementioned lower alkyl groups which are further substituted with one or more halogen atoms described above, preferably xe2x80x94CF3.
The term xe2x80x9cnitrogen-containing saturated heterocyclic groupxe2x80x9d throughout the specification refers to a 5- to 7-membered heterocyclic group containing one or two nitrogen atoms on the ring, which may further contain one O or S atom and may form a bridge structure or may be fused with one benzene ring. Preferred examples of such heterocyclic group are pyrrolidinyl, piperazinyl, piperidyl and morpholinyl. Preferred examples of the nitrogen-containing saturated heterocyclic group as xe2x80x94NR2R3 are 1-pyrrolidinyl, 1-piperazinyl, piperidino and morpholino, with particular preference to morpholino.
The term xe2x80x9carylxe2x80x9d is used throughout the specification to mean an aromatic cyclic hydrocarbon group. An aryl having 6 to 14 carbon atoms is preferable. Preferred examples of such aryl are phenyl and naphthyl.
The term xe2x80x9cheteroarylxe2x80x9d refers to a 5- or 6-membered monocyclic heteroaryl containing 1 to 4 hetero atoms selected from N, S and O as well as a bicyclic heteroaryl fused to a benzene ring. The heteroaryl may be partially saturated. Preferred examples of the monocyclic heteroaryl are furyl, thienyl, pyrrolyl, imidazolyl, pyrazolyl, thiazolyl, isothiazolyl, oxazolyl, triazolyl, tetrazolyl, pyridyl, pyrimidinyl, pyridazinyl and pyrazinyl. Examples of the bicyclic heteroaryl are preferably benzofuranyl, benzothienyl, benzothiadiazolyl, benzothiazolyl, benzimidazolyl, indolyl, isoindolyl, indazolyl, quinolyl, isoquinolyl, cinolinyl, quinazolinyl, quinoxalinyl and benzodioxolyl. Specific examples of the partially saturated heteroaryl are 1,2,3,4-tetrahydroquinolyl, etc. Particularly preferred are 5- to 6-membered monocyclic groups, more preferably imidazolyl, thiazolyl, triazolyl, pyridyl and pyrazinyl.
Examples of a xe2x80x9c5- or 6-membered monocyclic heteroaryl containing 1 or 2 hetero atoms selected from O, S and Nxe2x80x9d in B include a furan, thiophene, pyrole, imidazole, pyrazole, thiazole, isothiazole, oxazole, pyridine, pyrimidine, pyridazine and pyrazine ring. Preferably, it is a pyridine, pyrazine or thiophene ring. More preferable, it is a pyridine ring.
The substituents for the xe2x80x9caryl which may have one or more substituentsxe2x80x9d, xe2x80x9cheteroaryl which may have one or more substituentsxe2x80x9d, xe2x80x9ccycloalkyl which may have one or more substituentsxe2x80x9d, xe2x80x9ccycloalkenyl which may have one or more substituentsxe2x80x9d or xe2x80x9cnitrogen-containing saturated heterocyclic group which may have one or more substituentsxe2x80x9d are 1xcx9c5 substituents, which may be the same or different. Preferably, these substituents are selected from Group A described below. Each of R, Rxe2x80x2 and Rxe2x80x3, which may be the same or different, represents H or a lower alkyl (the same shall apply hereinafter).
Group A: -a lower alkyl, -a lower alkenyl, -a lower alkynyl, -a halogen, -a halogenated lower alkyl, -a lower alkylene-OR, xe2x80x94NO2, xe2x80x94CN, xe2x95x90O, xe2x80x94OR, xe2x80x94O a halogenated lower alkyl, xe2x80x94O-a lower alkylene-NRRxe2x80x2, xe2x80x94O-a lower alkylene-OR, xe2x80x94O-a lower alkylene-an aryl, xe2x80x94SR, xe2x80x94SO2-a lower alkyl, xe2x80x94SO-a lower alkyl, xe2x80x94COOR, xe2x80x94COO-a lower alkylene-an aryl, xe2x80x94COR, xe2x80x94CO-an aryl, -an aryl, xe2x80x94CONRRxe2x80x2, xe2x80x94SO2NRRxe2x80x2, xe2x80x94NRRxe2x80x2, xe2x80x94NRxe2x80x3-a lower alkylene-NRRxe2x80x2, xe2x80x94NRxe2x80x2-a lower alkylene-OR, xe2x80x94NR-a lower alkylene-an aryl, xe2x80x94NRCO-a lower alkyl, xe2x80x94NRSO2-a lower alkyl, -a cycloalkyl and -a cycloalkenyl.
When R4, R4a and R4b represent xe2x80x9can aryl which may have one or more substituentsxe2x80x9d or xe2x80x9ca heteroaryl which may have one or more substituentsxe2x80x9d, the substituents are 1 to 5 groups selected from a) through c) below, which may be the same or different.
a): -a lower alkyl, -a lower alkenyl, -a lower alkynyl, -a halogen, -a halogenated lower alkyl, -a lower alkylene-OR, xe2x80x94NO2, xe2x80x94CN, xe2x95x90O, xe2x80x94O-halogenated lower alkyl, xe2x80x94SO2-a lower alkyl, xe2x80x94SO-a lower alkyl, xe2x80x94COOR, xe2x80x94COO-a lower alkylene-an aryl, xe2x80x94COR, xe2x80x94CO-an aryl, xe2x80x94CONRRxe2x80x2, xe2x80x94SO2NRRxe2x80x2, -Cyc or -Alp-Cyc (wherein Alp represents a lower alkylene, a lower alkenylene or a lower alkynylene, and Cyc represents an aryl which may have 1 to 5 substituents selected from Group A, a heteroaryl which may have 1 to 5 substituents selected from Group A, a nitrogen-containing saturated heterocyclic group which may have 1 to 5 substituents selected from Group A, a cycloalkyl which may have 1 to 5 substituents selected from Group A, or a cycloalkenyl which may have 1 to 5 substituents selected from Group A; the same shall apply hereinafter).
b): xe2x80x94NR-E-F (wherein E represents xe2x80x94COxe2x80x94, xe2x80x94COOxe2x80x94, xe2x80x94CONRxe2x80x2xe2x80x94, xe2x80x94SO2NRxe2x80x2xe2x80x94 or xe2x80x94SO2-; F represents -Cyc or -(a lower alkyl, a lower alkenyl or a lower alkynyl which may be substituted by one or more substituents selected from the group comprising of -a halogen, xe2x80x94NO2, xe2x80x94CN, xe2x80x94OR, xe2x80x94O-a lower alkylene-NRRxe2x80x2, xe2x80x94O-a lower alkylene-OR, xe2x80x94SR, xe2x80x94SO2-a lower alkyl, xe2x80x94SO-a lower alkyl, xe2x80x94COOR, xe2x80x94COR, xe2x80x94CO-an aryl, xe2x80x94CONRRxe2x80x2, xe2x80x94SO2NRRxe2x80x2, xe2x80x94NRCO-a lower alkyl, xe2x80x94NRRxe2x80x2, xe2x80x94NRxe2x80x2-a lower alkylene-OR, xe2x80x94NRxe2x80x3-a lower alkylene-NRRxe2x80x2 and -Cyc); and the same shall apply hereinafter).
c): -Z-Rxe2x80x2, -Z-Cyc, -Z-Alp-Cyc, -Z-Alp-Zxe2x80x2-Rxe2x80x2 or -Z-Alp-Zxe2x80x2-Cyc (wherein each of Z and Zxe2x80x2, which may be the same or different, independently represents O, S or NR; and the same shall apply hereinafter).
The particularly preferred ones are -a lower alkylene-OR, xe2x80x94CONRRxe2x80x2, xe2x80x94NR-CO-Cyc1 (wherein Cyc1 is -an aryl which may have 1xcx9c5 substituents selected from Group A, -a heteroaryl which may have 1xcx9c5 substituents selected from Group A, or -a nitrogen- containing saturated heterocyclic group which may have 1xcx9c5 substituents selected from Group A, and the same applies hereinbelow), xe2x80x94NRxe2x80x94SO2-Cyc1, xe2x80x94OR, xe2x80x94NRRxe2x80x2, xe2x80x94O-a lower alkylene-NRRxe2x80x2 and xe2x80x94O-a lower alkylene-(a nitrogen-containing saturated ring which may have 1xcx9c5 substituents selected from Group A).
When n is 2 to 4, each R1 group may be the same or different, independently.
In the compounds which are shown by formulas (I), (Ia) and (Ib) of the present invention, the following compounds are preferred:
(1) Compounds in which R2 and R3 forms xe2x80x94NR2R3 which is a nitrogen-containing saturated heterocyclic group which may have 1xcx9c2 substituents selected from the group comprising of xe2x80x94OH, xe2x95x90O and -a lower alkyl;
(2) Compounds in which R2 and R3 forms xe2x80x94NR2R3 which is -morpholino;
(3) Compounds in which W is N;
(4) Compounds in which R4, R4a or R4b represents -(an aryl which may have one or more substituents) or -(a heteroaryl which may have one or more substituents);
(5) Compounds in which B represents a benzene ring; R1 represents -a lower alkyl, -a lower alkenyl, -a lower alkynyl, -a cycloalkyl, -an aryl which may have one or more substituents, -a heteroaryl which may have one or more substituents, -a halogen, xe2x80x94NO2, xe2x80x94CN, -a halogenated lower alkyl, xe2x80x94ORb, xe2x80x94SRb, xe2x80x94SO2-Rb, xe2x80x94SOxe2x80x94Rb, xe2x80x94COORb, xe2x80x94COxe2x80x94Rb, xe2x80x94CONRaRb, xe2x80x94SO2NRaRb, xe2x80x94NRaRb, xe2x80x94NRaxe2x80x94CORb, xe2x80x94NRaxe2x80x94SO2Rb, xe2x80x94Oxe2x80x94COxe2x80x94NRaRb or xe2x80x94NRaCOxe2x80x94COORb;
(6) Compounds in which B is a pyridine, pyrazine or thiophene ring and n is 0;
(7) Compounds in which X represents N, Y represents O and n is 0; and
(8) Compounds in which R4, R4a or R4b represents an aryl which has one or more substituents selected from the group comprising of -a lower alkylene-OR, xe2x80x94CONRRxe2x80x2, xe2x80x94NRxe2x80x94CO-Cyc1, xe2x80x94NRxe2x80x94SO2-Cyc1, xe2x80x94OR, xe2x80x94NRRxe2x80x2, xe2x80x94O-a lower alkylene-NRRxe2x80x2 and xe2x80x94O-a lower alkylene- (a nitrogen-containing saturated heterocyclic group which may have 1xcx9c5 substituents selected from Group A).
The particularly preferred compounds shown by general formula (Ia) are those having R4a which is a phenyl having at least one substituent which is selected from of the group comprising of xe2x80x94OH, xe2x80x94NH2, xe2x80x94NH-a lower alkyl, xe2x80x94N(a lower alkyl)2, xe2x80x94O-a lower alkylene-NH2 and xe2x80x94O-a lower alkylene-(a nitrogen-containing saturated heterocyclic group which may be substituted by a lower alkyl).
Moreover, the following compounds shown by general formula (Ib) are particularly preferred:
(1) Compounds in which W represents N, R4b represents -(an aryl which may have one or more substituents), and R2 and R3 form xe2x80x94NR2R3 which is -morpholino;
(2) Compounds in which B represents a benzene ring, n is 1 or 2, and R1 represents -a halogen, xe2x80x94NO2, xe2x80x94CN, -a halogenated lower alkyl, xe2x80x94ORb, xe2x80x94SRb, xe2x80x94NRaRb, xe2x80x94NRaxe2x80x94CORb or xe2x80x94NRaxe2x80x94SO2Rb; and
(3) Compounds in which B represents a pyridine, pyrazine or thiophene ring, n is 0, and R4b represents a phenyl which has at least one substituent which is selected from xe2x80x94OH, xe2x80x94CH2OH and xe2x80x94CONH2.
Among the compounds of the present invention, the preferred ones which are shown by general formula (Ia) are (Co 17) 6-amino-3xe2x80x2-(4-morpholinopyrido[3xe2x80x2,2xe2x80x2:4,5]furo[3,2-d]pyrimidin-2-yl)nicotinanilide, (Co 33) 4-(4-morpholinopyrido[3xe2x80x2,2xe2x80x2:4,5]furo[3,2-d]pyrimidin-2-yl)aniline, (Co 50) 3-(4-morpholinopyrido[3xe2x80x2,2xe2x80x2:4,5]furo[3,2-d]pyrimidin-2-yl)phenol, (Co 69) 4-morpholino-2-[3-(2-piperazin-1-ylethoxy)phenyl]pyrido[3xe2x80x2,2xe2x80x2:4,5]furo[3,2-d]pyrimidine, (Co 73) 3xe2x80x2-(4-morpholinopyrido[3xe2x80x2,2xe2x80x2:4,5]furo[3,2-d]pyrimidin-2-yl)acrylanilide, and salts thereof. The preferred ones which are shown by general formula (Ib) are (Co 144) N-[2-(3-benzenesulfonylaminophenyl)-4-morphoniloquinazolin-6-yl]acetamide, (Co 164) 3-(4-morpholinopyrido[4,3-d]pyrimidin-2-yl)phenol, (Co 172) 3-(4-morpholinopyrido[3,2-d]pyrimidin-2-yl)phenol, (Co 174) 3-(4-morpholinopyrido[3,4-d]pyrimidin-2-yl)phenol, (Co 186) 3-(6-methoxy-4-morpholinoquinazolin-2-yl)phenol, (Co 190) 3-(4-morpholinothieno[3,2-d]pyrimidin-2-yl)phenol, (Co 191) 3-(4-morpholinopteridin-2-yl)phenol, and salts thereof.
The compound of this invention may exist in the form of geometrical isomers or tautomers depending on the kinds of substituent groups, and these isomers in separated forms or mixtures thereof are included in the present invention. Also, the compound of the present invention may have asymmetric carbon atoms, so that optical isomer forms may exist based on such carbon atoms. All of the mixtures and the isolated forms of these optical isomers are included in the present invention.
Some of the compounds of the invention may form salts. There is no particular limitation so long as the formed salts are pharmacologically acceptable. Specific examples of acid salts are salts with inorganic acids such as hydrochloric acid, hydrobromic acid, hydroiodic acid, sulfuric acid, nitric acid, phosphoric acid, etc., organic acids such as formic acid, acetic acid, propionic acid, oxalic acid, malonic acid, succinic acid, fumaric acid, maleic acid, lactic acid, malic acid, tartaric acid, citric acid, methanesulfonic acid, ethanesulfonic acid, aspartic acid, glutamic acid, etc. Specific examples of basic salts include salts with inorganic bases containing metals such as sodium, potassium, magnesium, calcium, aluminum, etc., or salts with organic bases such as methylamine, ethylamine, ethanolamine, lysine, ornithine, etc. In addition, various hydrates and solvates and polymorphism of the compound (I), (Ia) or (Ib) and salts thereof are also included in this invention.
(Processes for producing compounds)
Hereinafter representative processes for producing the compounds of the present invention are described below. In these processes, functional groups present in the starting materials or intermediates may be suitably protected with protective groups, depending upon the kinds of functional groups. In view of the preparation technique, it may be advantageous to protect the functional groups with groups that can be readily reverted to the original functional groups. When required, the protective groups are removed to give the desired products. Examples of such functional groups are amino, hydroxy, carboxyl, etc. Examples of the protective groups which may be used to protect these functional groups are shown in, e.g., Greene and Wuts, xe2x80x9cProtective Groups in Organic Synthesisxe2x80x9d, second edition. These protective groups may be appropriately employed depending upon reaction conditions.

(Here and hereinafter, L represents a leaving group.)
This process for producing the compounds (I) of the present invention comprises converting the compounds shown by general formula (II) to reactive derivatives thereof (III) in a conventional manner and then reacting an amine (IV) with the reactive derivatives. When another reactive site containing the leaving group L also exists on the ring A or the substituent R4 in the reactive derivatives (III), the same or different amine (IV) may be reacted again, if necessary. In a similar manner, when the A ring or R4 of the compounds of the present invention has a leaving group L such as a chloro or fluoro, transformations of functional groups may be conducted such as a hydrolysis reaction according to a method described in J. Am. Chem. Soc., 68, 1288 (1946) or an ipso-substitution reaction using alkoxide as a reacting agent according to a method described in Tetrahedron Lett., 40, 675 (1999).
The leaving group shown by L is preferably a halogen, or an organic sulfonyloxy group, e.g., methanesulfonyloxy, p-toluenesulfonyloxy, etc.
The reaction for preparing the reactive derivatives (III) can be carried out by the usual procedures. Where the leaving group is chlorine, phosphorus oxychloride, oxalyl chloride or thionyl chloride can be reacted under cooling or heating or at room temperature in an inert organic solvent or without. As such an inert organic solvent, there is an aromatic hydrocarbon solvent such as benzene or toluene; an ethereal solvent such as tetrahydrofitran (THF), 1,4-dioxane, etc.; a halogenated hydrocarbon solvent such as dichloromethane, chloroform, etc.; and a basic solvent such as pyridine or collidine. These solvents may be used alone or as a mixture of two or more. The solvent is optionally selected depending on the kinds of starting compounds. The addition of a base (preferably a dialkylaniline, triethylamine, ammonia, lutidine, collidine, etc.), phosphorus chloride (e.g., phosphorus pentachloride), a quaternary ammonium salt (e.g., tetraethylammonium chloride), or an N,N-dialkylamide compound (e.g., dimethylformamide (DMF)) may be advantageous in some cases from the viewpoint of accelerating the reaction. Where the leaving group is sulfonyloxy, the active intermediates (III) can be synthesized from the corresponding sulfonyl chloride by the usual procedures, e.g., using a method described in Tetrahedron Lett. 23 (22), 2253 (1982) or Tetrahedron Lett. 27 (34), 4047 (1986).
The reaction for producing the compounds (I) from the reactive derivatives (III) and the amine (IV) can be carried out by reacting the amine (IV) in an inert organic solvent or in the absence of any solvents under cooling or heating or at room temperature. The solvent described above is available and it may be used singly or as a mixture of two or more. The addition of an inorganic base such as sodium hydride, or an organic base such as triethylamine (TEA), pyridine or 2,6-lutidine, may be advantageous in some cases from the viewpoint of accelerating the reaction

(Wherein Rd is a lower alkyl which may have one or more substituents and Rb has the same definition as defined above; and the same shall apply hereinafter.)
This process comprises O-alkylation of the hydroxy-substituted compounds shown by general formula (Ia) or (Ic) in a conventional manner to obtain the compounds (Ib) or (Id). The reaction may be carried out, e.g., by reacting the compounds (Ia) or (Ic) with an alkylating agent such as an alkyl halide or a sulfonic acid ester in the presence of a base such as triethylamine, potassium carbonate, sodium carbonate, cesium carbonate, sodium hydroxide, sodium hydride or potassium t-butoxide. The reaction temperature can be under cooling or heating or at room temperature, and can be appropriately chosen depending on the kinds of starting compounds. When water is used or contained as a solvent in an O-alkylation reaction, the reaction may be accelerated by the addition of a phase transfer catalyst such as tetra n-butylammonium hydrogensulfate.
Another method for the O-alkylation reaction is Mitsunobu reaction. For example, methods described in Synthesis, 1 (1981) or modified methods may be used. For the hydroxyethylation of a hydroxyl group, methods using carbonate ester such as [1,3]dioxolane-2-one are also effective. As an example, methods described in J. Am. Chem. Soc., 68, 781 (1946) can be used.
Moreover, when functional groups exist on Rb and Rd of the compounds (Ib) and (Id) of the present invention, known reactions may be employed to convert the functional group. For example, when a hydroxyl group is present on Rb and Rd, the aforementioned O-alkylation reaction can be conducted, and when a leaving group is present such as a halogen, an appropriate alcohol or amine can be reacted with utilizing the conditions of said O-alkylation or N-alkylation described hereinafter in Production Method 4. When an ester group is present, the functional group can be converted to a carboxylic acid, hydroxymethyl group, and amido, using a method described hereinafter in Production Method 3.
The starting compounds (Ia) and (Ic) used in this process can be prepared by the method described for Production Method 1, using starting compounds whose OH group has been protected by an acyl type protective group (e.g., acetyl or tosyl). Further, when phosphorus oxychloride is used as a reacting agent for synthesizing reactive derivatives (III) and then a desired amino is reacted to synthesize (I), protective groups for OH group may be removed and O-phosphoramides may be produced, depending on the kind of starting compounds, a protective group, reaction conditions and conditions for work-up. In that case, for example, using a method described in Chem. Pharm. Bull.,37, 2564 (1989), phosphoramides groups can be removed. Other general protective groups can be introduced and removed by the methods described in xe2x80x9cProtective Groups in Organic Synthesisxe2x80x9d supra.

(Wherein Rf is a lower alkyl and Rg is a lower alkyl which may have one or more substituents; and the same shall apply hereinafter.)
Production Method 3 comprises transformations of the functional groups of the ester compounds of the present invention shown by general formula (Ie) to produce the hydroxymethyl compounds (If), carboxylic acid derivatives (Ig) and amide derivatives (Ih) of the present invention, respectively. Each of the reactions can be carried out in a conventional manner, e.g., as described in Jikken Kagaku Kouza (Encyclopedia for Experimental Chemistry) edited by Nihon Kagaku Kai (Japanese Association of Chemistry) and published by Maruzen Co., Ltd., and xe2x80x9cProtective Groups in Organic Synthesisxe2x80x9d supra.
Preferably, the reduction to give the hydroxymethyl compounds (If) can be conducted in an inert organic solvent to the reactions, e.g., an ethereal solvent or an aromatic hydrocarbon solvent, using a reducing agent such as lithium aluminum hydride, lithium borohydride, zinc borohydride, boran, Vitride, etc. The hydrolysis to give the carboxylic acid derivatives (Ik) can be conducted by reacting with lithium hydroxide, sodium hydroxide or potassium hydroxide in a single solvent selected from methanol, ethanol, THF and water, or a mixture of two or more. The amidation to give the amide compounds (Ih) may be performed by converting carboxylic acids to reactive derivatives such as acyl halides (acyl chlorides, etc.) or acid anhydrides, and then reacting the reactive derivatives with amines. In the reaction with amines, it is preferred to conduct the reaction in an inert organic solvent in the presence of a base (an inorganic base such as sodium hydroxide, or an organic base such as TEA, diisopropylethylamine or pyridine). Furthermore, the amidation using the carboxylic acid as a starting compound can also be carried out in an inert organic solvent in the presence of a condensation agent such as (1-ethyl-3-(3-dimethylaminopropyl) carbodiimide (EDCI), 1,1xe2x80x2-carbonylbis-1H-imidazole (CDI), etc.). In this case, an additive such as 1-hydroxybenzotriazol (HOBt) or the like may also be added to the reaction. The reaction temperature and solvent can be appropriately chosen depending on the kinds or the like of starting compounds.

(Wherein Rxe2x80x2 has the same definition as defined above, Rh is -a lower alkyl which may have one or more substituents, Ri is -Cyc or -Alp which may have one or more substituents, a C ring is a nitrogen-containing saturated heterocyclic group which may have one or more substituents, and Rj is xe2x80x94H, -a lower alkyl, -an aryl, etc.; and the same shall apply hereinafter.)
Production Method 4 comprises the reduction of the nitro compounds shown by general formula (Ii) to the corresponding amino compounds (Ij) and then subjecting the amino compounds (Ij) to various modification reactions including N-alkylation, amidation, sulfonamidation, conversion to the corresponding urea, conversion to the corresponding carbamic acid, imidation or conversion to the corresponding thiazoles, to give the compounds (Ik), (Im), (In), (Io), (Ip), (Iq) and (Ir), respectively. These products can be appropriately subjected to further known modification reactions such as N-alkylation, if necessary.
These reactions can all be carried out in a conventional manner, e.g., using the methods described in xe2x80x9cJikken Kagaku Kouzaxe2x80x9d supra, or xe2x80x9cProtective Groups in Organic Synthesisxe2x80x9d supra. Preferred procedures in these methods are described below.
The reduction of the nitro compounds can be carried out in an alcoholic solvent such as methanol in a gaseous hydrogen atmosphere using palladium on carbon (Pdxe2x80x94C).
When various aldehydes are employed as the starting compounds, the N-alkylation can be conducted by reductive amination using aldehydes and reducing agents such as sodium borohydride, sodium triacetoxyborohydride or sodium cyanoborohydride. Reducting amination using Dean-Stark apparatus could be useful, too. When an alkyl halide such as methyl iodide or benzyl bromide, or dimethyl sulfate is employed as an alkylating agent, the reaction can be carried out in an inert organic solvent, e.g., DMF, acetonitrile or toluene, in the presence of base such as potassium carbonate, sodium hydroxide or sodium hydride, under cooling or heating or at room temperature. For monoalkylation, an example of useful procedure is as follows: protection of amino group by acyl group such as trifluoroacetyl, alkylation of acylamide by conventional methods using halogenated alkyl, and removal of protection. The dialkylation can be conducted by reacting 2 equivalents or more of a halogenated alkyl. For dimethylation, the reaction with formalin in formic acid at room temperature or under heating is also useful.
The amidation reaction may be performed in a similar manner to that described above for Production Method 3. The sulfonamidation can be carried out in an inert organic solvent using a reactive derivative such as an acid halide (acid chloride, etc.) or an acid anhydride. The conversion to the corresponding urea can be conducted by reacting with isocyanates in an inert organic solvent, e.g., an aromatic hydrocarbon solvent, under cooling or heating or at room temperature. The conversion to the corresponding carbamic acids can be conducted by reacting chloroformate derivatives in an inert organic solvent under cooling or heating or at room temperature. The imidation can be carried out using agents such as succinic anhydride or maleic anhydride.
The conversion to the corresponding aminothiazole compounds can be conducted by converting the amino compounds to the corresponding thiourea derivatives and then reacting the derivatives with an xcex1-halogenated ketone. Compounds (Ij) can be converted into the thiourea derivatives by methods described in, e.g., Synth. Commun. 1998, 28 (8), 1451; J. Org. Chem., 1984, 49 (6), 997, Org. Synth., 1963, IV, 180; J. Am. Chem. Soc., 1934, 56, 1408, etc. The conversion of the thiourea derivatives into the thiazole derivatives can be conducted by reacting the thiourea derivatives with the xcex1-halogenated ketone in an alcoholic solvent such as ethanol or a carbonyl solvent such as methyl ethyl ketone, under cooling or heating or at room temperature. The addition of a base (potassium carbonate, sodium carbonate, etc.) may be effective in some cases from the viewpoint of accelerating the reaction.

(Wherein Rk and Rm each represents -a lower alkyl which may have one or more substituents.)
Production Method 5 comprises converting the nitro compounds of the invention shown by general formula (Is) to the corresponding amino compounds (It) and then subjecting them to various modification reactions to obtain the other compounds of the present invention. Each reaction can be carried out as described for Production Method 4.
Other compounds included in the present invention can be obtained in the same manner as described above or by using methods well known to those skilled in the art. For instance, the reactions are carried out appropriately using methods described in xe2x80x9cJikken Kagaku Kouzaxe2x80x9d supra, or xe2x80x9cProtective Groups in Organic Synthesisxe2x80x9d supra.
For example, the demethylation reaction of compounds with an aryl group into the corresponding phenol derivatives can be carried out by the methods described in xe2x80x9cProtective Groups in Organic Synthesisxe2x80x9d supra, i.e., the method of reacting with a demethylating agent such as sodium cyanide or potassium cyanide in a solvent such as dimethylsulfoxide (DMSO), etc., at room temperature or under heating.
The starting compounds (II) for the synthesis of the present invention can be performed in conventional manners, e.g., by the reactions shown in the following synthetic routes.
Process 1
(Wherein Rn is a lower alkyl; and the same shall apply hereinafter.)
The starting compounds (IIc) can be synthesized by a cyclization reaction of amide intermediates (5) or cyclization conducted by reacting anthranylic acid derivatives (1) as the starting compounds with imidates (6). Conventional cyclization reactions for preparing pyrimidine ring are available for the cyclization reaction for this purpose. For instance, the method described in Chem. Pharm. Bull., 39 (2), 352 (1991) can be used for the cyclization of the intermediates (5) and the intermediates (1) and (6) as the starting materials can be cyclized by the method described in J. Med. Chem., 9, 408 (1966). The amide intermediates (5) can be synthesized by amidation of the aniline derivatives (4) in a conventional manner, or by sequential conversions of esterification of a carboxylic acid in (1), acylation of an amino, and amidation of the ester group according to conventional methods. For example, the amide intermediates (5) can be obtained in accordance with the methods described in J. Med. Chem., 33, 1722 (1990), Eur. J. Med. Chem.-Chim. Ther., 9(3), 305 (1974), etc. When (3) is obtained by acylation using (2) as the starting materials, diacylation may take place depending on the starting compounds and reaction conditions. In such a case, treatment with basic conditions will give desired monoacyl compounds (3).
Process 2
The starting compounds (IId) can be synthesized by cyclization of the amide intermediates (12) or by cyclization of the ester intermediates (7) and the amide compounds (10). The intermediates (12) can be cyclized in the same manner as described above; where the intermediates (7) and (10) are used as the starting compounds, the cyclization can be carried out by the method described in, e.g., J. Med. Chem., 37, 2106 (1994). The amide intermediates (12) can be prepared by conversion of the functional group in ester compounds (7) in a conventional manner. The bicyclic ester intermediates (9) can be synthesized by formation of a 5-membered ring by reacting nitrile compounds (7) with ester compounds (8) in the presence of a base, for example, in accordance with the method described in J. Org. Chem., 37, 3224 (1972) or J. Heterocycl. Chem., 11 (6), 975 (1974), etc.

The starting compounds (IIe) can be synthesized, e.g., by cyclization of the starting compounds (14). Preferably, the compounds (14) are heated in a solvent with a high boiling point such as diphenyl ether or in the absence of any solvents. The starting compounds (14) can be synthesized in a conventional manner, e.g., by. condensation of the corresponding anilines (13) with the compounds (15).
Each of the reaction products obtained by the aforementioned production methods is isolated and purified as a free base or a salt thereof. The salt can be produced by a usual salt forming method. The isolation and purification are carried out by employing usually used chemical techniques such as extraction, concentration, evaporation, crystallization, filtration, recrystallization, various types of chromatography and the like.
Various forms of isomers can be isolated by the usual procedures making use of physicochemical differences among isomers. For instance, racemic compounds can be separated by means of a conventional optical resolution method (e.g., by forming diastereomer salts with a conventional optically active acid such as tartaric acid, etc. and then optically resolving the salts) to give optically pure isomers. A mixture of diastereomers can be separated by conventional means, e.g., fractional crystallization or chromatography. In addition, an optical isomer can also be synthesized from an appropriate optically active starting compound.
The compounds of the present invention exhibit a PI3K inhibitory activity and therefore, can be utilized in order to inhibit abnormal cell growths in which PI3K plays a role. Thus, the compounds are effective in the treatment of disorders with which abnormal cell growth actions of PI3K are associated, such as restenosis, atherosclerosis, bone disorders, arthritis, diabetic retinopathy, psoriasis, benign prostatic hypertrophy, atherosclerosis, inflamation, angiogenesis, immunological disorders, pancreatitis, kidney disease, cancer, etc. In particular, the compounds of the present invention possess excellent cancer cell growth inhibiting effects and are effective in treating cancers, preferably all types of solid cancers and malignant lymphomas, and especially, leukemia, skin cancer, bladder cancer, breast cancer, uterus cancer, ovary cancer, prostate cancer, lung cancer, colon cancer, pancreas cancer, renal cancer, gastric cancer, brain tumor, etc.
The pharmacological effect of the compounds according to the invention have been verified by the following pharmacological tests.
Test Example 1 Inhibition of PI3K (p110xcex1 subtype)
Inhibition was determined using enzyme (bovine p110xcex1) prepared in the baculovirus expression system. Bovine p110 was prepared according to a modification from the method by I. Hiles et al., Cell, 70, 419 (1992). Each compound to be assayed was dissolved in DMSO and the obtained 10 mM DMSO solution was serially diluted with DMSO.
The compound (0.5 xcexcl) to be assayed and enzyme were mixed in 25 xcexcl of buffer solution (40 mM Tris-HCl (pH 7.4), 200 mM NaCl, 2 mM dithiothreitol, 5 mM MgCl2). Then, 25 xcexcl of 5 mM Tris-HCl (pH 7.4) buffered solution supplemented with 10 xcexcg PI (Sigma), 2xcexc Ci [xcex3-32P] ATP (Amersham Pharmacia) and 80 xcexcM non-radiolabeled ATP (Sigma) was added to the mixture to initiate the reaction. After reacting at 37xc2x0 C. for 15 minutes, 200 xcexcl of 1M HCl and 400 xcexcl of CHCl3/MeOH (1:1) were added to the reaction mixture. The resulting mixture was stirred and then centrifuged. After the organic layer was again extracted twice with 150 xcexcl of MeOH/1M HCl (1:1). The radioactivity was measured using Cerenkov light.
The IC50 inhibition activity was defined by a 50% inhibition concentration of each compound assayed, which was converted from the radioactivity determined as 100% when DMSO alone was added and as 0% when no enzyme was added.
The compounds of the prevent invention exhibited an excellent p110xcex1 subtype inhibition activity. For example, IC50 of Compound (hereinafter, abbreviated as Co) 10, Co 17, and Co 24 were less than 1 xcexcM.
Moreover, compounds of the prevent invention were confirmed to have inhibiting activities against other subtypes (such as a C2 xcex2 subtype).
Test Example 2Colon Cancer Cell Growth Inhibition
HCT116 cells from a colon cancer cell line were cultured in McCoy""s 5A medium (GIBCO) supplemented with 10% fetal bovine serum. HCT116 cells were inoculated on a 96 well plate (5000 cells/well) followed by overnight incubation. The test compound diluted with the medium was added to the medium in a final concentration of 0.1 to 30M (final DMSO concentration, 1%). After incubation over 72 hours, Alamar Blue reagent was added to the medium. Two hours after the addition, a ratio of fluorescent intensity at an excitation wavelength of 530 nm to that at an emission wavelength of 590 nm was measured to determine the IC50. Co 14, Co 24, Co 25, Co 31, Co 46 and Co 47 of the present invention exerted an excellent cancer cell growth inhibition activity.
Test Example 3Melanoma Cell Growth Inhibition
A375 cells from a melanoma cell line were cultured in DMEM medium (GIBCO) supplemented with 10% fetal bovine serum. A375 cells at 10,000 cells/100 xcexcl were added to a 96 well plate which contained 1 xcexcl/well of the test compounds (final concentration of 0.001xcx9c30 xcexcM). After incubation for over 46 hours, Alamar Blue reagent was added to the medium (10 xcexcl/well). Two hours after the addition, a ratio of fluorescent intensity at an excitation wavelength of 530 nm to that at an emission wavelength of 590 nm was measured to determine the IC50 of the test compounds in the same manner as in the above examples.
The compounds of the prevent invention exhibited an excellent cancer cell growth inhibition activity. For example, Co17, Co 33, Co 50, Co 69, Co 164, Co 172, Co 174, Co 186, Co 190 and Co 191 exerted a good melanoma cell growth inhibition activity. Their IC50 values were 0.33xcx9c4.26 xcexcM. Contrarily, the known PI3K inhibitor LY294002 showed a value of 8.39 xcexcM.
In addition to the above cancer cell lines, the compounds of the present invention exhibited excellent cancer cell growth inhibiting activities against Hela cells from a cervix cancer cell line, A549, H460 cells from a lung cancer cell line, COLO205, WiDr, Lovo cells from a colon cancer cell line, PC3, LNCap cells from a prostate cancer cell line, SKOV-3, OVCAR-3, CH1 cells from an ovary cancer cell line, U87 MG cells from a glioma cell line and BxPC-3 cells from a pancreas cancer cell line.
Test Example 4In vivo Cancer Cell Growth Inhibition
A single-cell suspension of HelaS3 (5xc3x97106 cells), a human cervix cancer cell line, was inoculated into the flank of female Balb/c nude mice by subcutaneously injection. When the tumor reached 100xcx9c200 mm3 in volume, test compounds were intraperitoneally administered once a day for two weeks. 20% Hydroxypropyl-xcex2-cyclodextrin/saline was intraperitoneally administered with the same schedule as a control group. The diameter of the tumors was measured with a vernier caliper at certain time intervals until one day after the final doze administration. The tumor volume was calculated by the following formula: xc2xdxc3x97(a shorter diameter)2xc3x97(a longer diameter).
In the present test, test compounds exhibited superior anti-tumor activities as compared with the control group.
The pharmaceutical composition of the present invention can be prepared in a conventional manner by mixing one or more compounds of the invention shown by general formula (I) with a carrier for medical use, a filler and other additives usually used in pharmaceutical preparation. The pharmaceutical composition of the invention may be administered either orally in the form of tablets, pills, capsules, granules, powders, liquid, etc., or parenterally such as by intravenous or intramuscular injection, in the form of suppositories, or through pernasal, permucosal or subcutaneous route.
For oral administration of the composition in the present invention, a solid composition in the form of, e.g., tablets, powders or granules is available. In such a solid composition, one or more active or effective ingredients are blended with at least one inert diluent such as lactose, mannitol, glucose, hydroxypropyl cellulose, microcrystalline cellulose, starch, polyvinyl pyrrolidone or magnesium aluminate metasilicate. The composition may further contain additives other than the inert diluent by the usual procedures. Examples of such additives include a lubricant such as magnesium stearate, a disintegrating agent such as calcium cellulose glycolate, a solubilization assisting agent such as glutamic acid or aspartic acid. Tablets or pills may be coated, if necessary, with films of sugar or a gastric or enteric substance such as sucrose, gelatin, hydroxypropyl cellulose, hydroxypropylmethyl cellulose phthalate, etc.
A liquid composition for oral administration includes pharmaceutically acceptable emulsions, solutions, suspensions, syrups, elixirs, etc. and contains an inert diluent conventionally employed, e.g., purified water or ethanol. In addition to the inert diluent above, the liquid composition may further contain an auxiliary agent such as a moistening agent or a suspending agent, a sweetener, a flavor and/or a preservative.
A composition for parenteral administration contains a sterile aqueous or non-aqueous solution, a suspension and an emulsion. Examples of the aqueous solution and suspension include distilled water for injection use and physiological saline. Typical examples of the non-aqueous solution and suspension are propylene glycol, polyethylene glycol, vegetable oil such as olive oil, an alcohol such as ethanol, polysorbate 80, and the like. These compositions may further contain a preservative, a moistening agent, an emulsifier, a dispersing agent, a stabilizer and a solubilization assisting agent. These compositions are sterilized, e.g., by filtering them through a bacteria retention filter, incorporating a bactericide or through irradiation. Alternatively, they may be prepared into a sterile solid composition, which is dissolved in sterile water or a sterile solvent for injection prior to use.
In the case of oral administration, suitable daily does is usually about 0.0001 to 50 mg/kg body weight, preferably about 0.001 to 10 mg/kg, more preferably about 0.01 to 1 mg/kg, and the daily does is administered once a day or divided into 2 to 4 doses per day. In the case of intravenous injection, suitable daily dose is usually about 0.0001 to 1 mg/kg body weight, preferably about 0.0001 to 0.1 mg/kg. And the daily does is administered once a day or divided into a plurality of doses per day. The dose may be appropriately determined for each case, depending on conditions, age, sex, etc.
The compounds of the present invention can be utilized alone, or in conjunction with other treatments (e.g., radiotherapy and surgery). Moreover, they can be utilized in conjunction with other antitumor agents, such as alkylation agents (cisplatin, carboplatin, etc.), antimetabolites (methotrexate, 5-FU, etc.), antitumor antibiotics (adriamymycin, bleomycin, etc.), antitumor vegetable alkaloids (taxol, etoposide, etc.), antitumor hormones (dexamethasone, tamoxifen, etc.), antitumor immunological agents (interferon xcex1, xcex2, xcex3, etc.), and so forth.